ABSTRACT
Objective To analyze the levels of cytokines (IL-2,IFN-γ,IL-6,IL-10) associated with Th1 and Th2 cells in HLA-DQ8 transgenic mice model of ocular experimental autoimmune myasthenia gravis (oEAMG) induced by recombinant H-AChR γ subunit immunization.Methods DQ8 mice were immunized with 20 μg of AChR γ subunit,20 μg of crude E. coli extract (E. coli group),or complete Freund's adjuvant (CFA) only (CFA group). All mice were immunized on days 0,30,and 60. Mice were euthanized 28 days after the third immunization,and draining lymph node cells (LNC) and spleen lymphocytes were cultured in vitro. The supernatant was collected to observe the interleukin(IL)-2,interferon(IFN)-γ,IL-6,IL-10 production by ELISA.Results LNCs and spleen lymphocytes of H-AChR γ subunit-immunized mice exhibited significantly enhanced IFN-γ (F=76.332,P<0.001;F=34.865,P<0.001) and IL-2 (F=42.835,P<0.001;F=38.030,P<0.001),which associated with Th1 cells,as compared to E. coli group and CFA group. There were no significant differences in IL-6 (F=1.325,P=0.284;F=1.935,P=0.166) and IL-10 (F=0.908,P=0.417;F=1.189,P=0.322) levels,which secreted by Th2 cells,among these three groups.Conclusion Th1 cytokines play key roles in the pathogenesis of oEAMG,while the mechanism of Th2 cytokines for oEAMG remains unclear.
Subject(s)
Animals , Mice , Cytokines , Escherichia coli , HLA-DQ Antigens , Interferon-gamma , Mice, Transgenic , Receptors, Cholinergic , Th1 Cells , Th2 CellsABSTRACT
Objective To explore the protective effect of neuregulin-1 (NRG-1) on heart function and myocardium in rats with sepsis and its mechanism . Methods Healthy male Sprague-Dawly (SD) rats were divided into three groups according to random number table method, with 6 rats in each group. Sepsis model was established by cecal ligation and puncture (CLP group); rats in sham operation group (sham group) underwent the same procedure except ligation. Rats in NRG-1 pre-treatment group (NRG group) were intravenously injected with recombinant human NGR-1 (rhNRG-1) at the dose of 10 μg/kg through tail vein; rats in CLP group and sham group were treated with the same amount of saline. At 24 hours after CLP, hemodynamic method was used to evaluate the cardiac function, and myocardial morphology was observed with hematoxylin and eosin (HE) staining, enzyme linked immunosorbent assay (ELISA) was used to detect the levels of cardiac troponin T (cTnT), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) in serum and macrophage migration inhibitor factor (MIF) in myocardial tissue. Results ① heart function: compared with the sham group, the mean arterial pressure (MAP), left ventricular systolic pressure (LVSP) and left ventricular pressure maximal rate of rise and fall (±dp/dt max) were significantly decreased in CLP group and NRG group, while the MAP, LVSP and ±dp/dt max in NRG group were significantly higher than those in CLP group [MAP (mmHg, 1 mmHg = 0.133 kPa): 125.78±8.15 vs. 113.05±5.85, LVSP (mmHg): 151.27±6.79 vs. 139.39±8.05, +dp/dt max (kPa/s): 4 389.59±332.38 vs. 3 706.85±451.31, -dp/dt max (kPa/s): 4 291.42±323.72 vs. 3 691.17±515.44, all 1 <0.05]. ②Myocardial injury: compared with the sham group, the levels of serum cTnT in CLP group and NRG group were significantly increased, while the levels of serum cTnT in NRG group were significantly lower than those in CLP group (ng/L: 206.37±67.28 vs. 344.13±80.95, 1 < 0.05), and the HE staining showed that myocardial pathological changes in NRG group were improved compared with the CLP group. ③Inflammatory mediators level: compared with the sham group, the levels of serum TNF-α, IL-1β and myocardial MIF were significantly increased in CLP group and NRG group, while the indicators in NRG group were lower than those in CLP group [TNF-α(ng/L): 52.77±3.43 vs. 97.19±13.98, IL-1β (ng/L): 40.25±5.48 vs. 56.05±6.88, MIF (μg/L): 1.92±0.16 vs. 2.87±0.10, all 1 <0.05]. Conclusion NRG-1 can reduce circulating levels of inflammatory factors in rats with sepsis, adjust myocardial MIF level, and alleviate myocardial cell injury, thereby improving cardiac function, and play a role in myocardial protection.
ABSTRACT
Objective To observe the effects of neuregulin‐1 (NRG‐1)on calcium transients in mouse ventricular myo‐cytes.Methods Nine C57BL/6 mice of male were randomly dived into 3 groups :the blank control group ,NRG‐1 group ,and iso‐proterenol(ISO)group.Their hearts were removed and immediately cannulated via the aorta and retrogradely perfused with en‐zymatic isolation solution to get single ventricular cell by Langendorff system.Myocytes were loaded with the Ca2+ indicator Fluo‐4 and subjected to electrical field stimulation at 0.5 or 1 Hz by using living cells workstation.The change of fluorescene in‐tensity was recorded simultaneously in each group.Results Compared with the blank control group ,NRG‐1 group had signifi‐cantly increased Ca2+ transient amplitude ΔF/F0 (n=10 ,P0.05).Conclusion NRG‐1 can increase Ca2+ transient amplitude and Ca2+ transients and reduce the time interval of Ca2+ transients.
ABSTRACT
Objective To investigate the sleep condition and its influence factors in old patients with hypertension. Methods A total of 100 old patients with hypertension were investigated by Pitts-burgh sleep quality index (PSQI) and serf-designed questionnaire of influence factors. Results 76 per-cents of the old patients with hypertension had sleep disorders.The total score of PSQI,sleep quality,time take to fall asleep,hours of sleep,sleep effficiency,sleep disorders,hypnotic drugs and every score of daytime dysfunction were significantly higher than domestic norm(P<0.01).Surroundings(strange circumstances or noisiness), physio-pathology (disease distress or high frequency of going to toilet during nighttime),psycho-logical situations(the fear to the disease or worrying about medical fees) and drags(use of diuretics or the adverse effects of some antihypertensive agents) were the important factors affecting sleep quality. Con-clusions Old patients with hypertension have poor sleep quality,which relates with multiple factors.Valid nursing interventions should be adopted in order to control the various factors which affect sleep quality to improve patients' sleep quality.
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the expression of myeloid differentiation protein-2 (MD-2) in the human endothelial cells and its role in lipopolysaccharide (LPS)-induced NF-kappaB activation in endothelial cells.</p><p><b>METHODS</b>In vitro cultured human umbilical vein endothelial cells (HUVEC) were employed in the study. The expression of MD-2 mRNA and protein, and the effect of LPS on the expression of its mRNA and protein were assessed with RT-PCR and Western blotting. The role of MD-2 in LPS-induced NF-kappaB activation and IL-8 production were investigated with gene transfection of mutant MD-2 cDNA (0.5, 1.0, 2.0 microg), pEF-BOS vacant vector (2.0 microg) and MD-2 plasmid (2.0 microg) into HUVEC, respectively.</p><p><b>RESULTS</b>There was MD-2 mRNA and protein expression in HUVECs before LPS stimulation, and it could be obviously upregulated by LPS in time and dose-dependent manner (MD-2 protein absorbency was 25 196 +/- 1 723 without LPS stimulation, which was obviously lower than that stimulated with 0.01 mg/L LPS (58 817 +/- 3 241, P < 0.01) for 6 hours. Transfection of mutant MD-2 cDNA could remarkably inhibit LPS-induced NF-kappaB activation and IL-8 production in endothelial cells.</p><p><b>CONCLUSION</b>MD-2 might play an important role in the LPS-induced NF-kappaB activation in HUVECs.</p>
Subject(s)
Humans , Cell Differentiation , Cells, Cultured , Endothelial Cells , Metabolism , Interleukin-6 , Genetics , Interleukin-8 , Metabolism , Lipopolysaccharides , Pharmacology , NF-kappa B , Metabolism , Umbilical Veins , Cell BiologyABSTRACT
<p><b>OBJECTIVE</b>To construct the multi-probe ribonuclease protection assay (RPA) template set to be used for detecting expression patterns of MD-2, TLR4, CD14 mRNAs in human peripheral blood mononuclear cells.</p><p><b>METHODS</b>The designed cDNA fragments of the three genes were generated by polymerase chain reaction (PCR) using specific primers and directionally cloned into EcoR I and Hind III sites of expression plasmid pSP72 containing the T7 promoter, the linearized plasmids was used as template to synthesize anti-sense RNA probes. Then we extracted total RNA from peripheral blood mononuclear cells (PBMC) and detected the dynamic expression patterns of the three genes with RPA method.</p><p><b>RESULTS</b>The proper sequence and orientation of the template set were confirmed by sequencing and the template set was successfully used to assay TLR4, MD-2 and CD14 mRNAs in human PBMC. The results showed that the three detected genes decreased transiently 1-3 hours after 100 ng/ml LPS stimulation.</p><p><b>CONCLUSIONS</b>These new RPA multi-probe set provided valuable tool for the simultaneous quantitative determination of expression of TLR4, CD14 and MD-2 mRNAs in both constitutive and inducible types.</p>